Our overall goal is to develop a urine DNA-based screening test for the early detection of colorectal cancer (CRC) that is noninvasive, patient-friendly, and more sensitive than existing noninvasive screening tests. Despite the availability of colonoscopy, the current standard of care for CRC screening, CRC remains the nation's second leading cause of cancer mortality because of the low compliance (<40%) due to inconvenience, fear of discomfort, and the risk involved in the invasive screening test. The 5-year survival rate for CRC is 93% if it is diagnosed at stage I but only 8% if diagnosed at stage IV [1, 2]. Thus, the need is urgent to increase CRC screening for more than 70 million people in the United States age 50 and older. Current available noninvasive CRC screening tests include the fecal occult blood test (FOBT), fecal immunochemical test (FIT), the stool DNA test (PreGenPlus, Exact Sciences, Maynard, MA), next generation stool DNA test, and the Septin 9 plasma test. Unfortunately, the high cost, the inconvenience of sample collection, or the low sensitivities of these tests result in an overall less than satisfactory compliance rate (~60%) for CRC screening. Therefore, a less unpleasant, noninvasive, low cost, and highly sensitive screening test is needed to enhance the compliance rate and to increase the rate of early detection of advanced adenoma and CRC to improve the prognosis of the disease. Urine contains nucleic acids that can be used for the early detection of diseases and cancers that occur at non-urinary tract sites. We have successfully detected mVIM in urine of patients with CRC with a sensitivity of 75% in a small pilot study. Based on these data, our mVIM urine test has been included in the EDRN 6000-subject phase II validation study (Protocol ID 320). JBS Science Inc. has performed preliminary experiments and proposes to develop a JBS CRC urine test detecting three markers, mVIM and mutated K-ras and BRAF DNA, to ensure that at least 70% sensitivity to detect CRC can be obtained in a blinded prevalidation study. JBS Science Inc has demonstrated the feasibility of several key areas of this proposal. The goal of this application will be to develop robust assays for K-ras and BRAF mutations combined with a developed mVIM assay for the JBS CRC urine test (Aim 1) and to train this test in an open-label training set and then validate this test in a blinded test set of urine samples (Aim 2) in the phase I study In phase II, we will further develop and evaluate the urine DNA test in a large validation study. PUBLIC HEALTH RELEVANCE: There is an urgent need to develop a sensitive, patient-friendly, noninvasive screening test to identify individuals who have high likelihood of having CRC and bring them to colonoscopy for confirmation and treatment, so CRC can be detected earlier and the prognosis of the disease can be improved. The goal of this phase I project is to explore the ability of a urine DNA test to detect early stages of colon cancer by analyzing colon cancer-associated genetic and epigenetic modifications.